Abstract
The quantitation of liberated reducing sugars by the copper-bicinchoninic acid (BCA) assay provides a highly sensitive method for the measurement of glycoside hydrolase (GH) activity, particularly on soluble polysaccharide substrates. Here, we describe a straightforward method adapted to low-volume polymerase chain reaction (PCR) tubes which enables the rapid, parallel determination of GH kinetics in applications ranging from initial activity screening and assay optimization, to precise Michaelis–Menten analysis.
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Acknowledgments
Funding from the Natural Sciences and Engineering Research Council of Canada (NSERC) via the Strategic Partnership Grants for Networks (for the Industrial Biocatalysis Network) and Discovery Grant programs is gratefully acknowledged. Equipment infrastructure was funded by the Canada Foundation for Innovation and the British Columbia Knowledge Development Fund. We thank Sean McDonald (Brumer group, UBC) for comments on an early version of this chapter.
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Arnal, G., Attia, M.A., Asohan, J., Brumer, H. (2017). A Low-Volume, Parallel Copper-Bicinchoninic Acid (BCA) Assay for Glycoside Hydrolases. In: Abbott, D., Lammerts van Bueren, A. (eds) Protein-Carbohydrate Interactions. Methods in Molecular Biology, vol 1588. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6899-2_1
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DOI: https://doi.org/10.1007/978-1-4939-6899-2_1
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